LOCUS       Exported                6179 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses improved CRY2PHR homo-oligomerization.
ACCESSION   .
VERSION     .
KEYWORDS    mCherry-CRY2clust
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6179)
  AUTHORS   Park H, Kim NY, Lee S, Kim N, Kim J, Heo WD
  TITLE     Optogenetic protein clustering through fluorescent protein tagging 
            and extension of CRY2.
  JOURNAL   Nat Commun. 2017 Jun 23;8(1):30. doi: 10.1038/s41467-017-00060-2.
  PUBMED    28646204
REFERENCE   2  (bases 1 to 6179)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6179
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      644..653
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             650..1354
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELY"
     primer_bind     complement(795..813)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1063..1082
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3004..3125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3041..3060)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3095..3114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3132..3587)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3269..3290)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3481..3500
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3614..3718
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3720..4077
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3743..3763)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3928..4063
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3990..4009
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4112..4906
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4166..4185)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4776..4795
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5094..5113)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5138..5185
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5514..6102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6003..6022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa
      661 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
      721 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
      781 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
      841 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
      901 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
      961 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1021 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1081 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1141 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     1201 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     1261 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     1321 ccgccactcc accggcggca tggacgagct gtacatccgg aaaccggtcg ccaccatgaa
     1381 gatggacaaa aagaccatcg tctggtttcg gagagatttg agaatagaag ataatcccgc
     1441 gctcgccgcc gcggcccacg agggttccgt cttccccgtt ttcatttggt gtcctgaaga
     1501 agaaggccag ttttatcccg gaagggcctc taggtggtgg atgaagcaaa gtctggccca
     1561 tcttagccag tcactgaaag cactgggcag tgatcttacc ctgatcaaga cacacaatac
     1621 catctctgcc atcctcgact gcatcagggt gaccggcgca acgaaagtcg tgtttaacca
     1681 cctgtacgat ccagttagtc tggtgcgcga ccacactgtg aaggagaagc tggtggaacg
     1741 ggggatcagt gtgcagagct acaacgggga ccttctgtac gagccatggg agatctattg
     1801 cgagaaaggg aaaccgttca cctccttcaa cagttactgg aagaaatgtt tggatatgtc
     1861 aatagagtcc gttatgttgc cccctccctg gagactgatg ccgattactg ctgctgcaga
     1921 ggccatctgg gcctgctcca tcgaggaact cggtctggaa aatgaagcag aaaagccaag
     1981 caatgcactt ctcactagag cctggagccc cggctggtct aatgccgaca agctgcttaa
     2041 cgagttcatc gaaaaacaac tgattgacta cgcgaagaac tccaagaaag tggtaggtaa
     2101 ctcaactagc ttgctctctc catatctcca ttttggcgag atttctgtcc gccatgtatt
     2161 tcagtgcgct cggatgaaac agattatctg ggctcgcgat aaaaacagcg aaggcgaaga
     2221 aagcgccgat ctgttcctgc gagggatcgg acttcgggaa tactcccggt atatatgttt
     2281 caactttcca ttcacacacg agcagagtct gttgtcccac ctcaggttct tcccctggga
     2341 cgccgatgtc gacaaattca aggcatggag acagggaagg acagggtacc cactcgtgga
     2401 tgctggcatg agagagctct gggctacagg ctggatgcac aaccgcatcc gggtaatcgt
     2461 gtcctcattt gctgtcaagt ttctgctcct gccttggaaa tggggaatga agtacttttg
     2521 ggataccctt ctcgacgccg acttggagtg tgacattctg ggatggcaat atattagcgg
     2581 gtcaattcct gacggccatg agttggacag gttggacaat ccggccttgc agggagctaa
     2641 gtatgatccc gaaggagagt atattcgaca gtggctcccc gagctggccc gacttcctac
     2701 ggagtggatt caccatcctt gggacgcacc actgacagtg ctcaaggcaa gcggggtgga
     2761 gctgggcacc aattacgcta agcctatagt tgatatagat acagcacgcg agctgctggc
     2821 taaagcgatc tctcgcactc gggaggcgca gattatgatc ggtgctgccg cccgggatcc
     2881 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     2941 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3001 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3061 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3121 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3181 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3241 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3301 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3361 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3421 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3481 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3541 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3601 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3661 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3721 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3901 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3961 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4021 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4081 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4141 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4201 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4261 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4321 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4381 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4441 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4501 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4561 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4621 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4681 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4741 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4801 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4861 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4921 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4981 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5041 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5101 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5161 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5221 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5281 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5341 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5401 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5461 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5521 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5581 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5641 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5701 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5761 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5821 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5881 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5941 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6001 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6061 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatc
//
